AC Electrothermal Flow Research Articles

Construction of on-site DNA pre-treatment device and rapid electrochemical biosensor set for Escherichia coli detection in milk

Restaurants and cafeterias are constantly exposed to bacterial contamination. In such environments, it is highly needed the rapid bacteria detection system with simple pre-treatment device. To solve this problem, this study presented the pre-treatment device and Rapid electrochemical (REC) biosensor set for detecting Escherichia coli (E. coli) gene in milk sample. The pre-treatment device is consisted of tetraethyl orthosilicate (TEOS)-coated bead that operated with sonication method, and it can effectively remove the E. coli residues and extract E. coli DNA well. By analyzing the sequences of various foodborne pathogens, we selected the highly variable region sequences of E. coli DNA to use as the bioprobe for REC biosensor. The selected thiol-modified bioprobe was immobilized onto the fabricated biosensor electrode. For detecting the extracted E. coli gene fragment rapidly, the alternating current electrothermal flow (ACEF) technique was introduced to REC-biosensor. ACEF technique minimize the target-bioprobe binding time to 10 mins. Using the extracted E. coli DNA fragments with the pre-treatment device, the proposed REC-biosensor shows the limit of detection (LOD) in milk samples was 9.235 × 10-5 ng/μL, and the target detection accuracy was 92.5 %, with a maximal error rate up to 6.730 %. Moreover, the biosensor demonstrated high selectivity in electrochemical performance evaluation (Matrix effect = 7.425 %). The developed pre-treatment device provides simple and quick preprocessing time (within 20 min) of E. coli, and REC biosensor provides the rapid detection of E. coli (within 10 min). In conclusion, the proposed pre-treatment device and biosensor set can be easily applied to other types of bacteria, virus or germs in the near future.

Establishment of the rapid electrochemical-SELEX monitoring system by using ACEF technique and its application to dengue virus aptasensor fabrication

Aptamers, which consist of single-stranded DNA or RNA, possess considerable potential as biomolecular recognition elements. The systematic evolution of ligands by exponential enrichment (SELEX) serves as a general method for synthesizing aptamers. However, the conventional SELEX process for monitoring aptamer-target binding is time-consuming and labor-intensive, highlighting the need for rapid monitoring and selection methods. To address this problem, this study introduces a novel approach called rapid electrochemical-SELEX (RE-SELEX) monitoring method, which significantly reduces the target-aptamer binding time from 3 h to 10 min. This process is achieved by employing an alternating current electrothermal flow (ACEF) technique. The Au micro gap-type electrode was used to RE-SELEX monitoring device. As a proof-concept of this method, the dengue virus (DENV) aptamers were synthesized. For confirming the feasibility of this DENV aptamer, we analyzed target binding of produced aptamers test including gel electrophoresis, dissociation constant analysis. Then, two aptamers were selected for constructing the rapid electrochemical DENV aptasensor. The RE biosensor was achieved by ACEF technique that reduce the target-aptamer binding within 10 min. The fabricated device can detect 37 fM of the target dengue envelope protein in diluted human serum well. Thus, the proposed method provides a reliable SELEX-monitoring method for the rapid synthesis of aptamers using a simple electrochemical platform.

AC electrothermal trapping for efficient nanoparticles enrichment in a microchannel

Trapping nanoparticles in a microfluidic device holds promise for enriching low-concentration species in samples. This article explores AC electrothermal (ACET) flow as an effective mechanism for nanoparticle trapping within a microfluidic system. Employing a precisely designed microfluidic device with coplanar symmetric electrodes, we systematically analyze the trapping performance of 100-nm polystyrene nanoparticles (PsNPs) dispersed in deionized water. Through fluorescence microscopy, we observe the dynamic behavior of PsNPs under the influence of an AC electric field. The concentration factor (CF) is introduced as a metric for evaluating trapping efficiency, revealing a remarkable up to 30-fold increase in concentration within the electrode gap. The quadratic relationship between electrothermal forces and electric field strength is investigated, highlighting the direct impact on trapping enhancement. By adjusting the applied voltage to vary the electric field strength, CF is found to significantly improve, reaching a peak value of 16.3 at 3.5 MV m−1. Furthermore, we explore the influence of frequency on ACET velocity, emphasizing the importance of aligning frequency and electric field strength for effective nanoparticle trapping. This study contributes valuable insights into the optimization of ACET for precise and controlled nanoparticle manipulation, holding promise for diverse applications in fields requiring nanoparticle enrichment.

A rapid field-ready electrical biosensor consisting of bismuthine-derived Au island decorated BiOCl nanosheets for Raphidiopsis raciborskii detection in freshwater.

Cyanobacteria play an essential role in nutrient cycling in aquatic ecosystems. However, certain species adversely affect the environment and human health by causing harmful cyanobacterial algal blooms (cyanoHABs) and producing cyanotoxins. To address this issue, continuous cyanoHAB monitoring has been considered; however, a gold standard has not yet been established. In this study, we aimed to develop a dual DNA-targeting capacitive-type biosensor for rapid field-ready monitoring of Raphidiopsis raciborskii, a causative species of cyanoHAB. To enhance the sensing signal, a plate-like Au-BiOCl nanocomposite was synthesized using a spontaneous carbonation process without additional additives. The alternating-current electrothermal flow (ACEF) technique was applied to enable rapid DNA and probe binding within 10 min. The limits of detection (LODs) for R. raciborskii RubisCO large subunit (rbcL) and RNA polymerase beta subunit (rpoB) genes diluted in deionized (DI) water were 4.89 × 10-17 and 3.89 × 10-17 M, respectively. Furthermore, the LODs of R. raciborskii rbcl and rpoB diluted in freshwater containing HAB were 2.55 × 10-16 and 3.84 × 10-16 M, respectively, demonstrating the field-ready applicability of the device. The fabricated cyanobacterial DNA-sensing platform enabled powerful species-specific detection using a small sample volume and low target concentration without a nucleic acid amplification step, dramatically reducing the detection time. This study has considerable implications for detecting HABs, early warning systems, and species-specific environmental monitoring technology.

Fast-response electrochemical biosensor based on a truncated aptamer and MXene heterolayer for West Nile virus detection in human serum

West Nile virus (WNV) is a mosquito-borne flavivirus that can cause West Nile fever, meningitis, encephalitis, and polio. Early detection of WNV is important to prevent infection spread on the field. To commercialize the electrochemical biosensor for WNV, rapid target detection with the cheap manufacture cost is essential. Here, we developed a fast-response electrochemical biosensor consisting of a truncated WNV aptamer/MXene (Ti3C2Tx) bilayer on round-type micro gap. To reduce the target binding time, the application of the alternating current electrothermal flow (ACEF) technology reduced the target detection time to within 10 min, providing a rapid biosensor platform. The MXene nanosheet improved electrochemical signal amplification, and the aptamer produced through systematic evolution of ligands by exponential enrichment process eliminated unnecessary base sequences via truncation and lowered the manufacturing cost. Under optimized conditions, the WNV limit of detection (LOD) and selectivity were measured using electrochemical measurement methods, including cyclic voltammetry and square wave voltammetry. The LOD was 2.57 pM for WNV diluted in deionized water and 1.06 pM for WNV diluted in 10% human serum. The fabricated electrochemical biosensor has high selectivity and allows rapid detection, suggesting the possibility of future application in the diagnosis of flaviviridae virus.

Synthesis of Isolated DNA Aptamer and Its Application of AC-Electrothermal Flow-Based Rapid Biosensor for the Detection of Dengue Virus in a Spiked Sample.

Dengue fever is an infectious disease caused by the dengue virus (DENV) and is transmitted by mosquitoes in tropical and subtropical regions. The early detection method at a low cost is essential. To address this, we synthesized the isolated DENV aptamer for fabricating a rapid electrochemical biosensor on a Au interdigitated microgap electrode (AuIMGE). The DENV aptamers were generated using the SELEX (systemic evolution of ligands by exponential enrichment) method for binding to DENV surface envelope proteins. To reduce the manufacturing cost, unnecessary nucleotide sequences were excluded from the isolation process of the DENV aptamer. To reduce the detection time, the alternating current electrothermal flow (ACEF) technique was applied to the fabricated biosensor, which can shorten the detection time to 10 min. The performance of the biosensor was evaluated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). In the diluted DENV protein solution, the linear range of the concentrations was from 1 pM to 1 μM and the LOD was 76.7 fM. Moreover, the proposed biosensor detected DENV in a diluted spiked sample at a linear range of 10-6 to 106 TCID50/mL, while the detection performance was proven with an LOD of 1.74 × 10-7 TCID50/mL along with high selectivity.

Rapid electrical biosensor consisting of DNA aptamer/carbon nanonetwork on microelectrode array for cardiac troponin I in human serum

Rapid detection of cardiac biomarkers is important for the early diagnosis of acute myocardial infarction (AMI). Currently, cardiac troponin I (cTnI) is the standard biomarker for precise diagnosis of AMI. It is essential for rapid diagnosis of AMI patient. Herein, we presented the rapid electrical biosensor composed of a carbon nanotube network (CNN) and DNA aptamer on a microelectrode array to detect cTnI in human serum sample. The island-shaped Au microgap electrodes array was used to provide a high signal-to-noise ratio response and maximize detection sensitivity. In addition, an alternating current electrothermal flow (ACEF) method was introduced to improve the affinity between the DNA aptamer and the target molecule, and cTnI was detected within 10 min using this method. The current change was analyzed using pulse measurement for 100 μs, and the integrated current method was proposed to improve the performance of the sensor platform. This biosensor can detect cTnI in a wide linear range from 1 pM to 100 nM in 10 % human serum. The limit of detection is 6.59 fM, indicating high selectivity. Thus, the proposed electrical biosensor platform may facilitate rapid and early detection of AMI in the future.

Fabrication of Rapid Electrical Pulse-Based Biosensor Consisting of Truncated DNA Aptamer for Zika Virus Envelope Protein Detection in Clinical Samples

Zika virus (ZV) infection causes fatal hemorrhagic fever. Most patients are unaware of their symptoms; therefore, a rapid diagnostic tool is required to detect ZV infection. To solve this problem, we developed a rapid electrical biosensor composed of a truncated DNA aptamer immobilized on an interdigitated gold micro-gap electrode and alternating current electrothermal flow (ACEF) technique. The truncated ZV aptamer (T-ZV apt) was prepared to reduce the manufacturing cost for biosensor fabrication, and it showed binding affinity similar to that of the original ZV aptamer. This pulse-voltammetry-based biosensor was composed of a T-ZV apt immobilized on an interdigitated micro-gap electrode. Atomic force microscopy was used to confirm the biosensor fabrication. In addition, the optimal biosensor performance conditions were investigated using pulse voltammetry. ACEF promoted aptamer-target binding, and the target virus envelope protein was detected in the diluted serum within 10 min. The biosensor waveform increased linearly as the concentration of the Zika envelope in the serum increased, and the detection limit was 90.1 pM. Our results suggest that the fabricated biosensor is a significant milestone for rapid virus detection.

Rapid electrochemical dual-target biosensor composed of an Aptamer/MXene hybrid on Au microgap electrodes for cytokines detection

Rapid detection methods for cytokine storm markers, such as tumor necrosis factor α (TNF-α) and interferon gamma (IFN-γ), are required. Herein, we describe the fabrication of a rapid electrochemical dual-target biosensor composed of aptamer/MXene (Ti3C2) nanosheet on an Au microgap electrode. Alternating current electrothermal flow (ACEF) significantly reduced the detection time (<10 min) to achieve the rapid biosensor construction. Additionally, MXene nanosheet was synthesized to improve the detection sensitivity. A dual-type Au microgap electrode was designed to measure TNF-α and IFN-γ levels using a single biosensor. Moreover, it performs 12 measurements using a small sample volume. To reduce detection time with stable aptamer-target complex formation, various ACEF conditions were evaluated and optimized to 10 min. Using the optimal conditions, the limit of detection (LOD) and selectivity were determined by electrochemical impedance spectroscopy (EIS). A linear region was observed in the concentration range of 1 pg/mL to 10 ng/mL of TNF-α and IFN-γ. The LOD of TNF-α and IFN-γ were 0.15 pg/mL and 0.12 pg/mL within 10 min, respectively. Furthermore, the proposed biosensor detected TNF-α and IFN-γ diluted in 10% human serum in the concentration range of 1 pg/mL to 10 ng/mL with LODs of 0.25 pg/mL and 0.26 pg/mL, respectively.

Rapid and Sensitive Detection of Nanomolecules by an AC Electrothermal Flow Facilitated Impedance Immunosensor.

Conventional immunosensors typically rely on passive diffusion dominated transport of analytes for binding reaction and hence, it is limited by low sensitivity and long detection times. We report a simple and efficient impedance sensing method that can be utilized to overcome both sensitivity and diffusion limitations of immunosensors. This method incorporates the structural advantage of nanorod-covered interdigitated electrodes and the microstirring effect of AC electrothermal flow (ACET) with impedance spectroscopy. ACET flow induced by a biased AC electric field can rapidly convect the analyte onto nanorod structured electrodes within a few seconds and enriches the number of binding molecules because of the excessive effective surface area. We performed numerical simulations to investigate the effect of ACET flow on the biosensor performance. The results indicated that AC bias to the side electrodes could induce fast convective flow, which facilitates the transport of the target molecules to the binding region located in the middle as a floating electrode. The temperature rise due to the Joule heating effect was measured using a thermoreflectance imaging method to find the optimum device operation conditions. The change of impedance caused by the receptors-target molecules binding at the sample/electrode interface was experimentally measured and quantified in real-time using the impedance spectroscopy technique. We observed that the impedance sensing method exhibited extremely fast response compared with those under no bias conditions. The measured impedance change can reach saturation in a minute. Compared to the conventional incubation method, the ACET flow enhanced method is faster in its reaction time, and the detection limit can be reduced to 1 ng/mL. In this work, we demonstrate that this sensor technology is promising and reliable for rapid, sensitive, and real-time monitoring of biomolecules in biologically relevant media such as blood, urine, and saliva.

A high-efficiency micromixing effect by pulsed AC electrothermal flow

PurposeThe on-chip high-throughput mixing process is one of the main challenges in the preparation process in clinical diagnostics. Because of high laminar flow in micro-channel, the fluid should be disturbed by external force. This paper aims to study pulsed AC electrothermal flow and the multiphysic interaction between the fluid behavior, external electric field, temperature field and convection-diffusion field to generate perturbation effect inside the channel.Design/methodology/approachA set of numerical simulations were carried out by multiphysic interactions between the fluid behavior, external electric field, temperature field and convection-diffusion field to generate the pulsed AC electrothermal flow inside the channel. Behavior of electrode–electrolyte system is discussed using the electrical lumped circuit model.FindingsHighly efficient temperature gradients are generated by applying pulsed electric potential over the electrodes; as a result, efficient secondary flows form inside the channel. The proposed method increases the interfacial contact area between the fluids and enhances the molecular diffusion transport phenomena. Maximum temperature rise of 4.1 K is observed in the gap between the electrodes for 0.08 S/m fluid medium, where the electric field is much stronger than elsewhere. Velocity field and concentration analysis reveal high performance perturbation effects for the mixing process. The periodic stretching and folding effects increase the interfacial contact area between the fluids by using pulsed AC electrothermal flow. Based on the results, 83 per cent mixing efficiency is achieved for 0.08 S/m fluid medium with a microchannel length of 400 µm. Both the mixing efficiency and generated temperature rise increase by increasing the fluid ionic strength.Originality/valueThe ability to generate low temperature rise is very important for AC electrothermally driven fluidic chips such as immunoassay chips. In the present research, a novel actuation mechanism has been proposed to generate AC electrothermal manipulation mechanism and enhance the mixing efficiency by using pulsed AC electrothermal flow.

An effective electrical sensing scheme using AC electrothermal flow on a biosensor platform based on a carbon nanotube network

We report a simple and efficient electrical sensing scheme that can be used to overcome the “diffusion limit” of affinity-based biosensors by incorporating the structural advantage of a concentric electrode biosensor platform and the microstirring effect of AC electrothermal flow (ACEF). To prove the effect of ACEF on the biosensor performance, we performed both simulations and experiments for the detection of cardiac troponin-I, which is a biomarker for acute myocardial infarction. The finite element simulation results indicate that AC bias to the electrode (which has a concentric structure in our device) can induce fast convection flow, which facilitates the transport of the target molecules to the binding region located between the two electrodes. In our device, the channel region made of a carbon nanotube network decorated with gold nanoparticles, which act as the attaching sites of the probe molecules, is used as a highly sensitive electrical channel. We find that the electrical sensing method exhibited extremely fast sensing speeds compared with those under no bias (diffusion-limited) conditions.

Enhanced model-based design of a high-throughput three dimensional micromixer driven by alternating-current electrothermal flow.

We propose a 3D microfluidic mixer based on the alternating current electrothermal (ACET) flow. The ACET vortex is produced by 3D electrodes embedded in the sidewall of the microchannel and is used to stir the fluidic sample throughout the entire channel depth. An optimized geometrical structure of the proposed 3D micromixer device is obtained based on the enhanced theoretical model of ACET flow and natural convection. We quantitatively analyze the flow field driven by the ACET, and a pattern of electrothermal microvortex is visualized by the micro-particle imaging velocimetry. Then, the mixing experiment is conducted using dye solutions with varying solution conductivities. Mixing efficiency can exceed 90% for electrolytes with 0.2 S/m (1 S/m) when the flow rate is 0.364 μL/min (0.728 μL/min) and the imposed peak-to-peak voltage is 52.5 V (35 V). A critical analysis of our micromixer in comparison with different mixer designs using a comparative mixing index is also performed. The ACET micromixer embedded with sidewall 3D electrodes can achieve a highly effective mixing performance and can generate high throughput in the continuous-flow condition.

Electrothermal flow on electrodes arrays at physiological conductivities

AC electrothermal (ET) flow is inevitable for microfluidic systems dissipating electric energy in a conducting medium. Therefore, many practical applications of biomicrofluidics are prone to ET flow. Here, a series of observations are reported on ET flow in a microfluidic chamber that houses three electrode pairs. The observations indicate that the variations in liquid conductivity and channel height critically impact the structure and magnitude of the flow field. Observations indicate that after a critical conductivity a global ET flow is present in the chamber, while at lower conductivities a vortex is present at every electrode edge. In addition, no ET flow is observed when the chamber height is kept below a critical value at physiological conductivity (∼1.5 S/m). The experimental observations are compared with the numerical simulations of ET flow. The validity of the assumptions made in the current AC ET flow theory is also discussed in the light of the experimental data. The observations can be critical while designing microfluidic systems that involve power dissipation in conductive fluids.

Long-range electrothermal fluid motion in microfluidic systems

AC electrothermal flow (ACEF) is the fluid motion created as a result of Joule heating induced temperature gradients. ACEF is capable of performing major microfluidic operations, such as pumping, mixing, concentration, separation and assay enhancement, and is effective in biological samples with a wide range of electrical conductivity. Here, we report long-range fluid motion induced by ACEF, which creates centimeter-scale vortices. The long-range fluid motion displays a strong voltage dependence and is suppressed in microchannels with a characteristic length below ∼300μm. An extended computational model of ACEF, which considers the effects of the density gradient and temperature-dependent parameters, is developed and compared experimentally by particle image velocimetry. The model captures the essence of ACEF in a wide range of channel dimensions and operating conditions. The combined experimental and computational study reveals the essential roles of buoyancy, temperature rise, and associated changes in material properties in the formation of the long-range fluid motion. Our results provide critical information for the design and modeling of ACEF based microfluidic systems toward various bioanalytical applications.

Optoelectric patterning: Effect of electrode material and thickness on laser-induced AC electrothermal flow.

Rapid electrokinetic patterning (REP) is an emerging optoelectric technique that takes advantage of laser-induced AC electrothermal flow and particle-electrode interactions to trap and translate particles. The electrothermal flow in REP is driven by the temperature rise induced by the laser absorption in the thin electrode layer. In previous REP applications 350-700 nm indium tin oxide (ITO) layers have been used as electrodes. In this study, we show that ITO is an inefficient electrode choice as more than 92% of the irradiated laser on the ITO electrodes is transmitted without absorption. Using theoretical, computational, and experimental approaches, we demonstrate that for a given laser power the temperature rise is controlled by both the electrode material and its thickness. A 25-nm thick Ti electrode creates an electrothermal flow of the same speed as a 700-nm thick ITO electrode while requiring only 14% of the laser power used by ITO. These results represent an important step in the design of low-cost portable REP systems by lowering the material cost and power consumption of the system.

Numerical simulation on the opto-electro-kinetic patterning for rapid concentration of particles in a microchannel.

This paper presents a mathematical model for laser-induced rapid electro-kinetic patterning (REP) to elucidate the mechanism for concentrating particles in a microchannel non-destructively and non-invasively. COMSOL(®)(v4.2a) multiphysics software was used to examine the effect of a variety of parameters on the focusing performance of the REP. A mathematical model of the REP was developed based on the AC electrothermal flow (ACET) equations, the dielectrophoresis (DEP) equation, the energy balance equation, the Navier-Stokes equation, and the concentration-distribution equation. The medium was assumed to be a diluted solute, and different electric potentials and laser illumination were applied to the desired place. Gold (Au) electrodes were used at the top and bottom of a microchannel. For model validation, the simulation results were compared with the experimental data. The results revealed the formation of a toroidal microvortex via the ACET effect, which was generated due to laser illumination and joule-heating in the area of interest. In addition, under some conditions, such as the frequency of AC, the DEP velocity, and the particle size, the ACET force enhances and compresses resulting in the concentration of particles. The conditions of the DEP velocity and the ACET velocity are presented in detail with a comparison of the experimental results.

Convection and mass transfer enhanced rapid capacitive serum immunoassay

The mechanism of conventional serum immunoassay typically relies on the nature of diffusion of the target analytes, which can cause slow specific binding and long detection times. This work presents a rapid capacitive immunoassay which enhances analyte transport by integrating the AC electrothermal (ACET) flow stirring and capacitive biosensing. The ACET flow conveys target molecules towards the probe-coated electrode surface, and leads to rapid preconcentration of target analytes and an instant capacitance change at the electrode–fluid interface. Numerical simulation of ACET convection and mass transfer is provided to analyze how the ACET flow affects the target molecule flux and binding with immobilized receptors. Bovine progesterone diagnosis is demonstrated as an experimental validation. Pregnancy positive and negative serum can be distinguished in 60 seconds. This rapid, sensitive label-free immunoassay can be readily integrated into many microsystems for the detection of many other immunoglobulins.

Fluid mixing using AC electrothermal flow on meandering electrodes in a microchannel

The mixing of fluids using AC electrothermal flow (AC-ETF) is presented. A pair of coplanar electrodes with a sinusoidal interelectrode gap was used to enhance the mixing in a microchannel. To demonstrate the performance of the mixer, conventional dilution experiments were conducted using Texas Red-labeled dextran. The dependence of mixing on the salt concentration (10(-3) ∼ 10(-1) mol dm(-3) ) of the solutions and frequency (100 kHz ∼ 5 MHz) of the applied voltage were investigated. AC-ETF was responsible for the mixing at salt concentrations >10(-2) mol dm(-3) , whereas the effect of AC-EOF was suggested to play a role at concentrations <10(-2) mol dm(-3) in the low-frequency region. The fluorogenic reaction of human serum albumin (HSA) with SYPRO Red in the mixer was also examined, and results showed that enrichment of fluorescence intensity and an almost uniform distribution of stained HSA were achieved. The present mixer can be employed as a powerful tool to facilitate efficient chemical and biomedical analysis on microfluidic devices.

G1-1-2 交流電場と温度勾配より誘起される流れ場の三次元計測(G1 マイクロ熱流体計測)

For transport-limited surface reaction, fluid flow transporting particles in a solution toward a reaction region is effective to enhance the reaction efficiency and rate. AC electrothermal (ET) flow, induced by property gradient due to temperature difference under electric field, is suited for the surface reaction enhancement. In this study, 3D flow field measurement was carried out to obtain ACET flow structure by micron-resolution particle image velocimetry (micro-PIV). A microfluidic device with an electrode having an inclined gap with 60 degrees respect to the streamwise direction was implemented. 3D velocity field is composed based on orthogonal 2D velocity fields by the observations of particle movement from side- and bottom-view. As a result, 3D flow of ACET was successfully measured. The results indicate that more effective flow concentrating particles would be available.